Application of CPNE1 serving as cervical squamous cell carcinoma treatment target and detection method

Abstract

The invention discloses application of CPNE1 serving as a therapeutic target of cervical squamous cell carcinoma and a detection method. According to the application of the CPNE1 in serving as the cervical squamous cell carcinoma treatment target and the detection method, Transwell detection is carried out on the development relation between the CPNE1 and cervical cancer, the expression level of the CPNE1 in a sample is measured, the measured expression level of the CPNE1 is compared with a preset normal range, whether CPNE1 expression is abnormal or not is determined on the basis of a comparison result, and therefore the development relation between the CPNE1 and the cervical cancer is determined. The expression of the CPNE1 in cervical cancer tissues is obviously increased, and the expression of the CPNE1 is further found to be related to clinical staging and prognosis of cervical cancer, so that early diagnosis molecular markers and treatment targets of cervical cancer are confirmed in later treatment, and therefore, directional target accurate treatment of advanced patients is ensured, and the clinical treatment cost is reduced. The problem that an effective diagnosis and treatment method for late metastatic or recurrent patients is lacked clinically in the existing screening technology is solved, and the later treatment effect of the patients is further improved.

Description

Technical Field

[0001] The present invention relates to the technical field of cell detection, and in particular to the use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and a detection method. Background Art

[0002] Cervical cancer is the most common gynecological malignancy, and the main histological type is squamous cell carcinoma, followed by adenocarcinoma. Cervical squamous cell carcinoma is divided into exophytic, endophytic, ulcerative, and endocervical types, and histologically they are well-differentiated, moderately differentiated, and poorly differentiated. With the widespread application of cervical cytology screening, cervical cancer and precancerous lesions can be discovered and treated early, and the incidence and mortality of cervical cancer have declined significantly; immunohistochemistry was used to detect the relationship between the expression of CPNE1 in cervical squamous cell carcinoma and clinical pathological characteristics; the effects of CPNE1 on the proliferation, cycle, apoptosis, migration, invasion, and tumorigenicity of cervical cancer cells in animals were detected, and the molecular mechanism of its promotion of cervical cancer progression was further explored.

[0003] Current screening technologies lack effective diagnostic and treatment methods for patients with advanced metastatic or recurrent disease, resulting in mediocre treatment outcomes for recurrent patients and a lack of targeted precision treatment for advanced patients. Therefore, it is urgent to find sensitive, efficient and specific molecular markers and therapeutic targets for early diagnosis of cervical cancer. Summary of the invention

[0004] 1. Technical issues to be solved

[0005] In view of the deficiencies in the prior art, the present invention provides a use and detection method of CPNE1 as a therapeutic target for cervical squamous cell carcinoma, performs Transwell detection on the developmental relationship between CPNE1 and cervical cancer, determines the expression level of CPNE1 in the sample, compares the determined CPNE1 expression level with a preset normal range, and determines whether the CPNE1 expression is abnormal based on the comparison result, thereby establishing the developmental relationship between CPNE1 and cervical cancer. The expression of CPNE1 in cervical cancer tissue is significantly increased, and it is further found that the expression of CPNE1 is related to the clinical stage and prognosis of cervical cancer, thereby confirming the early diagnostic molecular markers and therapeutic targets of cervical cancer in the later stage of treatment, thereby ensuring targeted precision treatment of advanced patients, and solving the problem that the current screening technology lacks effective diagnosis and treatment methods for advanced metastatic or recurrent patients in the clinic.

[0006] (II) Technical solution

[0007] To achieve the above object, the present invention provides the following technical solution: the use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and a detection method, comprising the following steps:

[0008] S1. Immunohistochemical detection: The sections were placed in 10 mM citrate buffer solution (pH = 6.0), heated in a pressure cooker for 10 min to repair the antigen, and after the temperature was cooled to 37 ° C, 3% hydrogen peroxide was added and incubated at 37 ° C for 10 min to block the activity of endogenous peroxidase. The sections were incubated with anti-CPNE1 monoclonal antibodies at 4 ° C for 24 hours, with a dilution ratio of 1:250. After being thoroughly rinsed with phosphate buffered saline (PBS), the sections were immunohistochemically stained with goat anti-mouse polymer labeled with horseradish peroxidase. Finally, the positive expression of CPNE1 protein was observed with the incubated color developer 3′-diaminobenzidine, and the cell nuclei were counterstained with hematoxylin;

[0009] S2. CCK8 detection:

[0010] (1) Prepare 100 μl of cell suspension in a 96-well plate. Pre-culture the plate in an incubator for 24 hours (37°C, 5% CO2);

[0011] (2) Add 10 μl of the test substance at different concentrations to the culture plate;

[0012] (3) Incubate the culture plate in an incubator for an appropriate period of time (24 and 48 hours);

[0013] (4) Add 10 μl of CCK8 solution to each well;

[0014] (5) Incubate the culture plate in an incubator for 1-4 hours;

[0015] (6) measuring the absorbance at 450 nm using an enzyme-labeled instrument;

[0016] S3. Cell streaking detection:

[0017] (1) Cell plating: Cells in the logarithmic growth phase are digested with trypsin to form a single cell suspension, and then inoculated into a 6-well culture plate. The density of cell plating should ensure that the fusion rate reaches 100% after overnight growth, and the final total volume of culture medium per well is 2 mL;

[0018] (2) Cell culture: Place the 6-well plate with cells in a 37°C, 5% CO2 incubator for 24 hours to ensure that the cells are fully attached and growing. Cell scratching: On the second day, use a 20 μL pipette tip to scratch perpendicularly to the black line on the back of the well plate so that the scratch intersects with the marked line. The purpose of this step is to scratch a clear mark on the cell monolayer to facilitate subsequent observation of cell migration.

[0019] (3) Washing cells: Wash the cells 2 to 3 times with sterile PBS to remove the scratched cells so that the remaining gaps can be clearly seen by the naked eye, and then replace with fresh serum-free or low-serum (<2%) culture medium;

[0020] (4) Cell culture and observation: Place the cells in a 37°C, 5% CO2 incubator and continue to culture them. After appropriate time points, such as 0, 6, 12, and 24 hours, remove the cells, observe them under a microscope, and take photos for record.

[0021] (5) Data analysis: Use ImageJ software to open the image, randomly draw 6 to 8 horizontal lines, and calculate the mean distance between cells to quantitatively analyze the migration ability of cells;

[0022] S4, Transwell assay:

[0023] (1) Starvation of cells: Before the experiment, the cells need to be starved to ensure that the cells are in the best state for the migration experiment;

[0024] (2) Glue laying: Dilute the matrix gel appropriately and spread it in the upper chamber. Allow it to polymerize overnight to provide a matrix for cell migration.

[0025] (3) Laying the culture medium in the lower chamber: Add culture medium containing 20% ​​FBS to the lower chamber to provide nutrients and growth factors for cell migration;

[0026] (4) Preparing cell suspension: Digest the cells with trypsin to prepare a cell suspension, and count the cells;

[0027] (5) Cell laying: The cell suspension is spread in the upper chamber and prepared to be placed in the incubator for culture;

[0028] (6) Continue culturing: Continue culturing in a cell culture incubator, usually for 24 or 48 hours, to allow the cells time to migrate;

[0029] (7) Wipe off the upper layer of cells: After the culture is completed, use a cotton swab to wipe off the cells and matrix gel in the upper chamber for observation and counting;

[0030] (8) Fixing cells: Use 4% paraformaldehyde to fix the cells for 20 minutes for subsequent staining and photography;

[0031] (9) Staining and photography: Use crystal violet or Giemsa staining and then take photos to record the migration of cells.

[0032] Preferably, a cervical tissue sample is obtained from the patient, and the expression level of CPNE1 in the sample is determined, and the determined CPNE1 expression level is compared with that of a normal control group, indicating that the patient may respond to treatment targeting CPNE1 based on the CPNE1 expression level being higher than a preset threshold of the normal control group.

[0033] Preferably, RNA is extracted from the obtained cervical tissue samples, and the relative expression level of CPNE1 mRNA is determined using real-time quantitative PCR technology. If the relative expression level of CPNE1 mRNA exceeds 5 times that of the normal control group, CPNE1 is considered to be highly expressed. Based on the relative expression level of CPNE1 mRNA exceeding 1.5 times that of the normal control group, it indicates that the patient may respond to treatment targeting CPNE1.

[0034] Preferably, the expression of CPNE1 protein in cervical tissue samples is detected by immunohistochemistry, and the average staining intensity of CPNE1 protein is calculated. If the average staining intensity is greater than or equal to 3, CPNE1 is judged to be highly expressed. Based on the average staining intensity of CPNE1 protein being greater than or equal to 3, it indicates that the patient may respond to treatment targeting CPNE1.

[0035] Preferably, flow cytometry is used to analyze CPNE1 protein expression in cervical tissue samples to determine the proportion of CPNE1-positive cells. If the proportion of CPNE1-positive cells exceeds 20%, CPNE1 is considered to be highly expressed. Based on the proportion of CPNE1-positive cells exceeding 20%, it indicates that the patient may respond to treatment targeting CPNE1.

[0036] Preferably, when the CPNE1 expression level is lower than 5 times the expression level in the normal control group, CPNE1 is judged to be low expression. If CPNE1 is low expressed, a treatment method targeting CPNE1 is not recommended. If CPNE1 is highly expressed, a treatment method targeting CPNE1 is recommended. Whether to use a treatment method targeting CPNE1 is determined based on whether the CPNE1 expression level is higher than 1.5 times the expression level in the normal control group.

[0037] Preferably, a cervical tissue sample is obtained from the patient, the expression level of CPNE1 in the sample is measured, the measured CPNE1 expression level is compared with a preset normal range, and whether the CPNE1 expression is abnormal is determined based on the comparison result.

[0038] (III) Beneficial effects

[0039] The present invention provides the use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and a detection method, which has the following beneficial effects:

[0040] (1) The use and detection method of CPNE1 as a therapeutic target for cervical squamous cell carcinoma. By performing Transwell detection on the developmental relationship between CPNE1 and cervical cancer, the expression level of CPNE1 in the sample is measured, and the measured CPNE1 expression level is compared with the preset normal range. Based on the comparison result, it is determined whether the CPNE1 expression is abnormal, thereby establishing the developmental relationship between CPNE1 and cervical cancer. The expression of CPNE1 in cervical cancer tissue is significantly increased. It is further found that the expression of CPNE1 is related to the clinical stage and prognosis of cervical cancer. Therefore, in the later treatment, the molecular markers and therapeutic targets for early diagnosis of cervical cancer are confirmed, thereby ensuring targeted precision treatment of advanced patients, solving the problem that the current screening technology lacks effective diagnosis and treatment methods for advanced metastatic or recurrent patients, and further improving the later treatment effect and cure rate of patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 The present invention shows that CPNE1 is highly expressed in cervical cancer tissues, wherein A is the HE image of cervical cancer tissues; B is the expression of CPNE1 in normal squamous epithelial tissues; C is the expression of CPNE1 in cancer tissues; D is the expression of CPNE1 in normal cervical squamous epithelial cells and cervical cancer cells;

[0042] Figure 2 The results of 24hCCK8 and 48hCCK8 flow cytometry detection of the present invention, where * indicates P < 0.05 compared with the NC group;

[0043] Figure 3 The numbers of cell streaking at 24h and 48h in the OE group of the present invention, where (A) is the 24h cell streaking graph, (B) is the 48h cell streaking graph, and (C) is the cell invasion detection result; # indicates P<0.05 compared with the NC group; DETAILED DESCRIPTION

[0044] Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without making any creative work shall fall within the scope of protection of the present invention.

[0045] The present invention provides a technical solution: the use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and a detection method, comprising the following steps:

[0046] S1. Immunohistochemical detection: The sections were placed in 10 mM citrate buffer solution (pH = 6.0), heated in a pressure cooker for 10 min to repair the antigen, and after the temperature was cooled to 37 ° C, 3% hydrogen peroxide was added and incubated at 37 ° C for 10 min to block the activity of endogenous peroxidase. The sections were incubated with anti-CPNE1 monoclonal antibodies at 4 ° C for 24 hours, with a dilution ratio of 1:250. After being thoroughly rinsed with phosphate buffered saline (PBS), the sections were immunohistochemically stained with goat anti-mouse polymer labeled with horseradish peroxidase. Finally, the positive expression of CPNE1 protein was observed with the incubated color developer 3′-diaminobenzidine, and the cell nuclei were counterstained with hematoxylin;

[0047] S2. CCK8 detection:

[0048] (1) Prepare 100 μl of cell suspension in a 96-well plate. Pre-culture the plate in an incubator for 24 hours (37°C, 5% CO2);

[0049] (2) Add 10 μl of the test substance at different concentrations to the culture plate;

[0050] (3) Incubate the culture plate in an incubator for an appropriate period of time (24 and 48 hours);

[0051] (4) Add 10 μl of CCK8 solution to each well;

[0052] (5) Incubate the culture plate in an incubator for 1-4 hours;

[0053] (6) measuring the absorbance at 450 nm using an enzyme-labeled instrument;

[0054] The results showed that the absorbance values ​​of the overexpression group at 24h and 481h were significantly higher than those of the control group and the empty load group, with statistical differences (P<0.05).

[0055] S3. Cell streaking detection:

[0056] (1) Cell plating: Cells in the logarithmic growth phase are digested with trypsin to form a single cell suspension, and then inoculated into a 6-well culture plate. The density of cell plating should ensure that the fusion rate reaches 100% after overnight growth, and the final total volume of culture medium per well is 2 mL;

[0057] (2) Cell culture: Place the 6-well plate with cells in a 37°C, 5% CO2 incubator for 24 hours to ensure that the cells are fully attached and growing. Cell scratching: On the second day, use a 20 μL pipette tip to scratch perpendicularly to the black line on the back of the well plate so that the scratch intersects with the marked line. The purpose of this step is to scratch a clear mark on the cell monolayer to facilitate subsequent observation of cell migration.

[0058] (3) Washing cells: Wash the cells 2 to 3 times with sterile PBS to remove the scratched cells so that the remaining gaps can be clearly seen by the naked eye, and then replace with fresh serum-free or low-serum (<2%) culture medium;

[0059] (4) Cell culture and observation: Place the cells in a 37°C, 5% CO2 incubator and continue to culture them. After appropriate time points, such as 0, 6, 12, and 24 hours, remove the cells, observe them under a microscope, and take photos for record.

[0060] (5) Data analysis: Use ImageJ software to open the image, randomly draw 6 to 8 horizontal lines, and calculate the mean distance between cells to quantitatively analyze the migration ability of cells;

[0061] S4, Transwell assay:

[0062] (1) Starvation of cells: Before the experiment, the cells need to be starved to ensure that the cells are in the best state for the migration experiment;

[0063] (2) Glue laying: Dilute the matrix gel appropriately and spread it in the upper chamber. Allow it to polymerize overnight to provide a matrix for cell migration.

[0064] (3) Laying the culture medium in the lower chamber: Add culture medium containing 20% ​​FBS to the lower chamber to provide nutrients and growth factors for cell migration;

[0065] (4) Preparing cell suspension: Digest the cells with trypsin to prepare a cell suspension, and count the cells;

[0066] (5) Cell laying: The cell suspension is spread in the upper chamber and prepared to be placed in the incubator for culture;

[0067] (6) Continue culturing: Continue culturing in a cell culture incubator, usually for 24 or 48 hours, to allow the cells time to migrate;

[0068] (7) Wipe off the upper layer of cells: After the culture is completed, use a cotton swab to wipe off the cells and matrix gel in the upper chamber for observation and counting;

[0069] (8) Fixing cells: Use 4% paraformaldehyde to fix the cells for 20 minutes for subsequent staining and photography;

[0070] (9) Staining and photography: Use crystal violet or Giemsa staining and then take photos to record the migration of cells.

[0071] The results showed that after lentiviral transfection and overexpression of CPNE1, the number of cell streaking in the OE group was higher than that in the NC group at 24h and 48h. Compared with the NC group, the number of invasive cells in the OE group was significantly increased (P<0.05).

[0072] Immunohistochemistry was used to detect the relationship between the expression of CPNE1 in cervical squamous cell carcinoma and clinical pathological characteristics; the effects of CPNE1 on the proliferation, cycle, apoptosis, migration, invasion and tumorigenicity of cervical cancer cells in animals were detected, and the molecular mechanism of its promotion of cervical cancer progression was further explored. Therefore, it is urgent to find sensitive, efficient and specific molecular markers and therapeutic targets for early diagnosis of cervical cancer. CPNE1 is the first member of the Copines family to be discovered and is related to the occurrence and development of tumors, but its relationship with cervical cancer is unknown. We found that the expression of CPNE1 in cervical cancer tissues was significantly increased, and further found that the expression of CPNE1 was related to the clinical stage and prognosis of cervical cancer. Therefore, we assumed that CPNE1 is an important molecule that promotes the progression of cervical cancer, and took cervical samples from patients for the following tests.

[0073] Cervical tissue samples were obtained from the patients, and the expression levels of CPNE1 in the samples were determined. The determined CPNE1 expression levels were compared with those of the normal control group. Based on the fact that the CPNE1 expression level was higher than the preset threshold of the normal control group, it was indicated that the patients might respond to the treatment targeting CPNE1. RNA was extracted from the obtained cervical tissue samples, and the relative expression of CPNE1 mRNA was determined by real-time quantitative PCR. If the relative expression of CPNE1 mRNA exceeded 5 times the expression of the normal control group, CPNE1 was considered to be highly expressed. Based on the fact that the relative expression of CPNE1 mRNA exceeded 1.5 times the expression of the normal control group, it was indicated that the patients might respond to the treatment targeting CPNE1. The expression of CPNE1 protein in cervical tissue samples was detected by immunohistochemistry, and the average staining intensity of CPNE1 protein was calculated. If the average staining intensity was greater than or equal to 3, CPNE1 was determined to be highly expressed. Based on the fact that the average staining intensity of CPNE1 protein was greater than or equal to 3, it was indicated that the patients might respond to the treatment targeting CPNE1. Respond to CPNE1 treatment. Flow cytometry is used to analyze CPNE1 protein expression in cervical tissue samples to determine the proportion of CPNE1-positive cells. If the proportion of CPNE1-positive cells exceeds 20%, CPNE1 is considered to be highly expressed. Based on the fact that the proportion of CPNE1-positive cells exceeds 20%, it indicates that the patient may respond to treatment targeting CPNE1. When the CPNE1 expression level is less than 5 times the expression level in the normal control group, CPNE1 is judged to be lowly expressed. If CPNE1 is lowly expressed, treatment methods targeting CPNE1 are not recommended. If CPNE1 is highly expressed, treatment methods targeting CPNE1 are recommended. Based on whether the CPNE1 expression level is higher than 1.5 times the expression level in the normal control group, it is decided whether to use treatment methods targeting CPNE1. A cervical tissue sample is obtained from the patient, and the expression level of CPNE1 in the sample is measured. The measured CPNE1 expression level is compared with the preset normal range. Based on the comparison result, it is determined whether CPNE1 expression is abnormal.

[0074] In summary, the use and detection method of CPNE1 as a therapeutic target for cervical squamous cell carcinoma, through Transwell detection of the developmental relationship between CPNE1 and cervical cancer, the expression level of CPNE1 in the sample is determined, and the determined CPNE1 expression level is compared with the preset normal range. Based on the comparison result, it is determined whether the CPNE1 expression is abnormal, thereby establishing the developmental relationship between CPNE1 and cervical cancer. The expression of CPNE1 in cervical cancer tissue is significantly increased, and it is further found that the expression of CPNE1 is related to the clinical stage and prognosis of cervical cancer. Therefore, in the later treatment, the early diagnostic molecular markers and therapeutic targets of cervical cancer are confirmed, thereby ensuring the targeted and precise treatment of advanced patients, and solving the problem that the current screening technology lacks effective diagnosis and treatment methods for advanced metastatic or recurrent patients in clinical practice.

[0075] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0076] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. Use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and a detection method, characterized in that: The following steps are included: S1. Immunohistochemical detection: The sections were placed in 10 mM citrate buffer solution (pH = 6.0), heated in a pressure cooker for 10 min to repair the antigen, and after the temperature was cooled to 37 ° C, 3% hydrogen peroxide was added and incubated at 37 ° C for 10 min to block the activity of endogenous peroxidase. The sections were incubated with anti-CPNE1 monoclonal antibodies at 4 ° C for 24 hours, with a dilution ratio of 1:

250. After being thoroughly rinsed with phosphate buffered saline (PBS), the sections were immunohistochemically stained with goat anti-mouse polymer labeled with horseradish peroxidase. Finally, the positive expression of CPNE1 protein was observed with the incubated color developer 3′-diaminobenzidine, and the cell nuclei were counterstained with hematoxylin; S2. CCK8 detection: (1) Prepare 100 μl of cell suspension in a 96-well plate. Pre-culture the plate in an incubator for 24 hours (37°C, 5% CO2); (2) Add 10 μl of the test substance at different concentrations to the culture plate; (3) Incubate the culture plate in an incubator for an appropriate period of time (24 and 48 hours); (4) Add 10 μl of CCK8 solution to each well; (5) Incubate the culture plate in an incubator for 1-4 hours; (6) measuring the absorbance at 450 nm using an enzyme-labeled instrument; S3. Cell streaking detection: (1) Cell plating: Cells in the logarithmic growth phase are digested with trypsin to form a single cell suspension, and then inoculated into a 6-well culture plate. The density of cell plating should ensure that the fusion rate reaches 100% after overnight growth, and the final total volume of culture medium per well is 2 mL; (2) Cell culture: Place the 6-well plate with cells in a 37°C, 5% CO2 incubator for 24 hours to ensure that the cells are fully attached and growing. Cell scratching: On the second day, use a 20 μL pipette tip to scratch perpendicularly to the black line on the back of the well plate so that the scratch intersects with the marked line. The purpose of this step is to scratch a clear mark on the cell monolayer to facilitate subsequent observation of cell migration. (3) Washing cells: Wash the cells 2 to 3 times with sterile PBS to remove the scratched cells so that the remaining gaps can be clearly seen by the naked eye, and then replace with fresh serum-free or low-serum (<2%) culture medium; (4) Cell culture and observation: Place the cells in a 37°C, 5% CO2 incubator and continue to culture them. After appropriate time points, such as 0, 6, 12, and 24 hours, remove the cells, observe them under a microscope, and take photos for record. (5) Data analysis: Use ImageJ software to open the image, randomly draw 6 to 8 horizontal lines, and calculate the mean distance between cells to quantitatively analyze the migration ability of cells; S4, Transwell assay: (1) Starvation of cells: Before the experiment, the cells need to be starved to ensure that the cells are in the best state for the migration experiment; (2) Glue laying: Dilute the matrix gel appropriately and spread it in the upper chamber. Allow it to polymerize overnight to provide a matrix for cell migration. (3) Laying the culture medium in the lower chamber: Add culture medium containing 20% ​​FBS to the lower chamber to provide nutrients and growth factors for cell migration; (4) Preparing cell suspension: Digest the cells with trypsin to prepare a cell suspension, and count the cells; (5) Cell laying: The cell suspension is spread in the upper chamber and prepared to be placed in the incubator for culture; (6) Continue culturing: Continue culturing in a cell culture incubator, usually for 24 or 48 hours, to allow the cells time to migrate; (7) Wipe off the upper layer of cells: After the culture is completed, use a cotton swab to wipe off the cells and matrix gel in the upper chamber for observation and counting; (8) Fixing cells: Use 4% paraformaldehyde to fix the cells for 20 minutes for subsequent staining and photography; (9) Staining and photography: Use crystal violet or Giemsa staining and then take photos to record the migration of cells.

2. The use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and the detection method according to claim 1, characterized in that: A cervical tissue sample is obtained from the patient, and the expression level of CPNE1 in the sample is measured. The measured CPIE1 expression level is compared with that of the normal control group. Based on the CPNE1 expression level being higher than the preset threshold of the normal control group, it is indicated that the patient may respond to treatment targeting CPIE1.

3. The use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and the detection method according to claim 1, characterized in that: RNA was extracted from the obtained cervical tissue samples, and the relative expression of CPNE1 mRNA was determined using real-time quantitative PCR technology. If the relative expression of CPNE1 mRNA exceeded 5 times that of the normal control group, CPNE1 was considered to be highly expressed. Based on the relative expression of CPNE1 mRNA exceeding 1.5 times that of the normal control group, it indicated that the patient may respond to treatment targeting CPNE1.

4. The use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and the detection method according to claim 1, characterized in that: The expression of CPNE1 protein in cervical tissue samples was detected by immunohistochemistry, and the average staining intensity of CPNE1 protein was calculated. If the average staining intensity was greater than or equal to 3, CPNE1 was judged to be highly expressed. Based on the average staining intensity of CPNE1 protein being greater than or equal to 3, it indicates that the patient may respond to treatment targeting CPNE1.

5. The use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and the detection method according to claim 1, characterized in that: CPNE1 protein expression in cervical tissue samples was analyzed using flow cytometry to determine the proportion of CPNE1-positive cells. If the proportion of CPNE1-positive cells exceeded 20%, CPNE1 was considered to be highly expressed. Based on the proportion of CPNE1-positive cells exceeding 20%, it indicated that the patient might respond to treatment targeting CPNE1.

6. The use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and the detection method according to claim 1, characterized in that: When the CPNE1 expression level is lower than 5 times the expression level in the normal control group, CPNE1 is judged to be low expression. If CPNE1 is low expressed, treatment methods targeting CPNE1 are not recommended. If CPNE1 is highly expressed, treatment methods targeting CPNE1 are recommended. Whether to use treatment methods targeting CPNE1 is determined based on whether the CPNE1 expression level is higher than 1.5 times the expression level in the normal control group.

7. The use of CPNE1 as a therapeutic target for cervical squamous cell carcinoma and the detection method according to claim 1, characterized in that: Obtain a cervical tissue sample from the patient, measure the expression level of CPNE1 in the sample, compare the measured CPNE1 expression level with a preset normal range, and determine whether CPNE1 expression is abnormal based on the comparison result.

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